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Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Published on: April 3, 2017
Single-cell transcriptomic analysis reveals characteristic feature of macrophage reprogramming in liver Mallory-Denk
Zixuan Fang1,2, Bei Zhong1, Yi Shi1,2
1The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China; The Qingyuan Affiliated Hospital of Guangzhou Medical University, Qingyuan People's hospital, Qingyuan, China.
Abstract:
Chronic liver diseases are highly linked with mitochondrial dysfunction and macrophage infiltration. Mallory-Denk bodies (MDBs) are protein aggregates associated with hepatic inflammation, and MDBs pathogenesis could be induced in mice by feeding 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). Here, we investigate the macrophage heterogeneity and the role of macrophage during MDBs pathogenesis on DDC-induced MDBs mouse model by single-nucleus RNA sequencing (snRNA-seq). We defined liver macrophages into four distinct subsets including monocyte-derived macrophages (MDMs) subset and three Kupffer cells (KCs) subsets (Gpnmbhigh KCs, Peam1high KCs, and Gpnmblow Pecam1low KCs). Particularly, we identified a novel Gpnmbhigh KCs subset as lipid-associated macrophage (LAM) with high expression of Trem2, CD63, and CD9. Interestingly, LAM showed a potential immunosuppressive characteristic by expressing anti-inflammatory genes IL-7R during the MDBs formation. Using contact and transwell co-culture systems, the released mtDNA from hepatocytes was found to induce the activation of inflammasome in macrophages. Furthermore, we revealed the damaged DNA could activate the NOD-like receptor family pyrin domain containing-3 (NLRP3) inflammasome and subsequently form apoptosis-associated speck-like protein containing a caspase recruit domain (ASC) specks of liver macrophages. Collectively, our results firstly revealed macrophage heterogeneity and inflammasome activation by mtDNA from injured liver during MDBs pathogenesis, providing crucial understanding of pathogenesis of chronic liver disease.
Insights
This study reveals macrophage heterogeneity in chronic liver disease, identifying a novel lipid-associated macrophage subset. Damaged mitochondrial DNA from injured liver cells activates inflammasomes in these macrophages during Mallory-Denk body formation.
Area of Science:
- Hepatology
- Immunology
- Cell Biology
Background:
- Chronic liver diseases involve mitochondrial dysfunction and macrophage infiltration.
- Mallory-Denk bodies (MDBs) are protein aggregates linked to hepatic inflammation.
- MDB pathogenesis can be modeled in mice using 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC).
Purpose of the Study:
- Investigate macrophage heterogeneity during MDB pathogenesis.
- Elucidate the role of macrophages in DDC-induced MDB mouse models.
- Understand inflammasome activation mechanisms in liver macrophages.
Main Methods:
- Single-nucleus RNA sequencing (snRNA-seq) on DDC-induced MDB mouse model.
- Macrophage subset identification and characterization.
- Co-culture systems to study hepatocyte-macrophage interactions and inflammasome activation.
Main Results:
- Defined four liver macrophage subsets: monocyte-derived macrophages (MDMs) and three Kupffer cell (KC) subsets (Gpnmbhigh KCs, Peam1high KCs, Gpnmblow Pecam1low KCs).
- Identified a novel Gpnmbhigh KC subset as lipid-associated macrophages (LAMs) expressing Trem2, CD63, CD9, and IL-7R, suggesting immunosuppressive properties.
- Demonstrated that released mitochondrial DNA (mtDNA) from injured hepatocytes activates the NOD-like receptor family pyrin domain containing-3 (NLRP3) inflammasome and ASC speck formation in liver macrophages.
Conclusions:
- Macrophage heterogeneity plays a significant role in MDB pathogenesis.
- Lipid-associated macrophages (LAMs) may exhibit immunosuppressive functions during MDB formation.
- mtDNA released from injured hepatocytes triggers inflammasome activation in liver macrophages, contributing to chronic liver disease pathogenesis.
